Putrescine or spermidine binding site of aminopropyltransferases and competitive inhibitors

Antagonistic Effect of N-(3-Aminopropyl)cyclohexylamine on Neurotrophic Action of Spermine in Primary Cultured Rat Hippocampal and Cerebellar Neurons

“…APCHA is supposed to work as a spermine synthase inhibitor by competing with spermidine at the active site of the enzyme (9). Because of the structural similarity between APCHA and spermine, APCHA may compete with spermine on the molecule that mediates the neurotrophic action of spermine.…”

“…The hydrochloride forms of APCHA and trans-4-methylcyclohexylamine (4MCHA) were synthetized at the Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Josai University, as described previously (9). Chemical structures of these analogues are shown in Fig.…”

“…N-(3-Aminopropyl)cyclohexylamine (APCHA) was originally developed as a potent and selective inhibitor of spermine synthase (9,10). To test if endogenous spermine biosynthesis contributes to neuronal survival and morphogenesis, we examined the effect of APCHA on cultured brain neurons.…”

Determination of Cellular Aminopropyltransferase Activity Using Precolumn Fluorescent Etheno-Derivatization with High-Performance Liquid Chromatography

“…21,22 Spdsyn activity assays for purified protein and cellular supernatant were carried out in a reaction buffer containing 0.1 M sodium phosphate pH 7.5, 0.1 mM dcSAM, 1 mM Put, 0.6 mM adenine, and 0.05 mM APCHA, which was added for cellular activity measurement to block Spmsyn, another endogenous Apt. 23 Each reaction mixture was incubated at 37 C for 1 h and the reaction was stopped by the addition of 1.1 M PCA, followed by centrifugation at 16000g for 20 min. Then, 1.5 M CAA and 0.2 M sodium acetate was added to the supernatant to adjust the pH to around 4.5, and the mixture was incubated for derivatization reaction at 70 C for 15 min.…”

Spermidine Regulates Insulin Synthesis and Cytoplasmic Ca2+ in Mouse Beta-TC6 Insulinoma Cells

“…However, since DFMO reduces the intracellular levels of polyamines including putrescine, spermidine and spermine, the functional role of each polyamine has not been fully elucidated. Meanwhile Shirahata et al (1991) developed two inhibitors, trans-4-methylcyclohexylamine (MCHA) and N-(3-aminopropyl)cyclohexylamine (APCHA), which are specific to spermidine synthase and spermine synthase, respectively. The effect of MCHA and APCHA has been investigated in rats (Shirahata et al, 1993;Kobayashi et al, 2006) and mice (Chu et al, 1994) in vivo, and cultured neurons (Chu et al, 1995), rat hepatoma HC11 cells (Beppu et al, 1995), mouse mammary carcinoma FM3A cells (Nishimura et al, 2005) and malaria parasite (Haider et al, 2005) in vitro.…”